The aim of this project is to identify reactions that are required for the initiation of Bacillus megaterium spore germination. For the current year of this project, the objectives were to determine the pathway of glucose metabolism during intitiation of germination, to further characterize mutants that were temperature-sensitive for initiation of germination and to isolate a mutant for leucine dehydrogenase. In addition, we wished to characterize the lytic phage (MP-7) isolated in this laboratory for B. megaterium. (a) From an analysis of (U-C14)-glucose metabolism, assays of 19 enzymes involved with glucose metabolism, isolation of mutants lacking either phosphofructokinase or 6-P-gluconate dehydrogenase and the ability of glucose analogues to initiate germination, it was concluded that glucose metabolism may not occur during the intital stages of germination. After initiation of germination, glucose is metabolized primarily by the Embden-Meyerhoff pathway. (b) Mutants that were temperature sensitive in any of the enzymes of glucose metabolism or the lytic enzyme activity. (c) Repeated attempts to isolate a leucine dehydrogenase mutant were negative. (d) Phage MP-7 was characterized in terms of properties of infection, host range and the DNA size and base composition. BIBLIOGRAPHIC REFERENCES: Vary, J.C. 1975. Properties of Bacillus megaterium temperature-sensitive germination mutants. J. Bacteriol. 121: 197-203. Hsieh, L.K. and J.C. Vary. 1975. Peptidoglycan hydrolysis during initiation of spore germination in Bacillus megaterium. In P. Gerhardt, H.L. Sadoff and R.N. Costilow (eds.). Spores VI. American Society for Microbiology, Washington, D.C.